Thursday 20th August

Lab Work

Digestion for check

by Audrey

BBa_K115017 (control +)
BBa_J23101 + GFP (control -)
BBa_K1707000 #2, #3 and #4
BBa_K1707013 #1
BBa_K1707030 #1
BBa_K1707019 #1
BBa_K1707020 #2
BBa_K1707021 #2
BBa_K1707027 #1
BBa_K1707035 #1
BBa_K1707036 #1

Mix for each tub:

- 2 µL plasmid
- 1 µL SpeI (Biolabs)
- 1 µL BspE1 (Biolabs)
- 2 µL Buffer 3 10x (Biolabs)
- 5 µL H2O

Electrophoresis to check PCR products

by Audrey

Agarose gel 1%, Migration 90V

Biobricks:

BBa_K1707031 #7 to #26

We can conclude that all clones aren't OK

Purification

by Pauline

BBa_K1707022 #1
BBa_K1707023 #1
BBa_K1707004 #2
BBa_K1707034 #1
BBa_K1707028 #1

With Macherey Nagel purification kit

Electrophoresis

by Pauline

Digestion products:

BBa_K115017 (control +)
BBa_J23101 + GFP (control -)
BBa_K1707000 #2, #3 and #4
BBa_K1707013 #1
BBa_K1707030 #1
BBa_K1707019 #1
BBa_K1707020 #2
BBa_K1707021 #2
BBa_K1707027 #1
BBa_K1707035 #1
BBa_K1707036 #1

Quantification:

BBa_K1707022 #1: ?
BBa_K1707023 #1: nothing
BBa_K1707004 #2: nothing
BBa_K1707034 #1: 20 µg/µL
BBa_K1707028 #1: 10 µg/µL

New culture

by Pauline

BBa_K1707004 #2

In 5mL LB + 5µL Ampicillin

Ligation

by Pauline

BBa_K1707037 = BBa_K1707028 + BBa_K1707034

Mix:

8 µL BBa_K1707028
10 µL BBa_K1707034
2 µL Ligase
2,5 µL Buffer
2,5 µL H2O

Incubation 4°C, ON

Quantification

by Pauline

BIobricks:

BBa_K1707022 #1
BBa_K1707023 #1

We can't see anything

Member present:

Instructors: Claire
Students: Pauline and Audrey

Back to the calendar

Team:Paris Saclay/Notebook/August/20

Contents

Thursday 20th August

Lab Work

Digestion for check

Electrophoresis to check PCR products

Purification

Electrophoresis

New culture

Ligation

Quantification